The present invention relates to a method of producing a corresponding xcex1-halo-xcex1,xcex2-saturated carbonyl compound from an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond by hydrogenating the xcex1,xcex2-carbon-carbon double bond using a microorganism belonging to the genus Acetobacter, Actinomyces, Acinetobacter, Agrobacterium, Aeromonas, Alcaligenes, Arthrobacter, Azotobacter, Bacillus, Brevibacterium, Burkholderia, Cellulomonas, Corynebacterium, Enterobacter, Enterococcus, Escherichia, Flavobacterium, Gluconobacter, Halobacteium, Halococccus, Klebsiella, Lactobacillus, Microbacterium, Micrococcus, Micropolyspora, Mycobacterium, Nocardia, Pseudomonas, Pseudonocardia, Rhodococcus, Rhodobacter, Serratia, Staphylococcus, Streptococcus, Streptomyces or Xanthomonas, preferably a microorganism belonging to the genus Pseudomonas or Burkholderia, more preferably Pseudomonas sp. SD810, Pseudomonas sp. SD811, Pseudomonas sp. SD812 or Burkholderia sp. SD816, or a microbial product thereof. The present invention also relates to novel microorganisms belonging to the genera Pseudomonas and Burkholderia, particularly Pseudomonas sp. SD810, Pseudomonas sp. SD811, Pseudomonas sp. SD812 and Burkholderia sp. SD816.
Furthermore, the present invention relates to a method of producing a corresponding xcex1-halo-xcex1,xcex2-saturated carbonyl compound as an S form compound with respect to the xcex1-position from an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond by hydrogenating the carbonxe2x80x94carbon double bond. This method can be used in the production of optically active carbonyl compounds such as various optically active (having an absolute S form configuration at the xcex1-position) saturated carboxylic acids or amides. The optically active carbonyl compounds are a highly valuable chiral building block which is difficult to prepare by classical chemical processes, and are materials useful particularly as a raw material of medical or agricultural chemicals.
In recent years, a method of producing various compounds, particularly optically active substances, by the reduction of a carbonxe2x80x94carbon double bond using a microorganism is drawing attention. To this effect, various methods of producing a corresponding xcex1,xcex2-saturated carbonyl compound having a substituent at the xcex1-position from a carbonyl compound having an xcex1,xcex2-carbon-carbon double bond and having a substituent at the xcex1-position by microbially reducing the carbonxe2x80x94carbon double bond have been reported (see, H. Simon, et al., Hoppe-Seyler""s Z. Physiol. Chem., 362, 33 (1981), H. Giesel, et al., Arch. Microbiol., 135, 51 (1983), H. G. W. Leuenberger, et al., Helv. Chim. Acta., 62, 455 (1979), R. Matsuno, et al., J. Ferm. Bioeng., 84, 195 (1997)). However, for example, according to the method of using bacteria as the microorganism, an anaerobe such as Clostridium kluyveri (DSM-555) or Clostridium sp. La-1 (DSM-1460) is used. Therefore, the growing rate of the microorganism is slow, it is difficult to increase the cell concentration and accordingly, the reaction rate is not satisfactorily high. Thus, these methods have a problem in profitability and operability.
The method using Clostridium theremosaccharolyticum disclosed in JP-A-63-003794 (the term xe2x80x9cJP-Axe2x80x9d as used herein means an xe2x80x9cunexamined published Japanese patent applicationxe2x80x9d) has an object of solving the above-descried problem by using a thermophilic bacterium. However, the bacterium used is still anaerobic, therefore, the growing rate and the reaction rate both are not satisfactorily high and the process involves use of hydrogen. Thus, the method fails in solving the problems in profitability and safety. Furthermore, the xcex1,xcex2-saturated carbonyl compound having a substituent at the xcex1-position produced by reducing a prochiral carbonyl compound having an xcex1,xcex2-carbon-carbon double bond and having a substituent at the xcex1-position using a microorganism is a compound having an absolute R form configuration and an S form configuration compound cannot be produced.
The method of reducing the xcex1,xcex2-carbon-carbon double bond using a bread yeast as the microorganism has general-purpose applicability because compounds over a wide range can be reduced. In addition, since the microorganism used is aerobic, good operability can be attained. Furthermore, the optically active substances produced include both S form and R form, therefore, this method is most abundant in the cases reported. However, the yeast grows slowly as compared with bacteria, the optical selectivity is not sufficiently high in many cases in the reduction reaction for obtaining a more optically active product, and reduction of an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond is not known.
As described above, in the technique of producing a corresponding xcex1-halo-xcex1,xcex2-saturated carbonyl compound from an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond by reducing the carbonxe2x80x94carbon double bond using a microorganism, a method satisfying all of the requirements regarding operability, profitability, safety and reaction properties is not yet known.
An object of the present invention is to provide a method of producing a corresponding xcex1-halo-xcex1,xcex2-saturated carbonyl compound from an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond by reducing the carbonxe2x80x94carbon double bond using a microorganism, which method can satisfy all of the requirements for operability, profitability, safety and reaction properties and ensure excellent optical selectivity.
As a result of thorough screening from soil, the present inventors have found that surprisingly, microorganisms capable of producing a corresponding xcex1-halo-xcex1,xcex2-saturated carbonyl compound from an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond by reducing the carbonxe2x80x94carbon double bond are distributed over a relatively wide genus range of the aerobes and facultative anaerobes. In particular, it has been found that strains having this activity are present in a large number in microorganisms belonging to the genera Pseudomonas and Burkholderia, and some of these strains can reduce an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond and thereby produce an extremely high-purity xcex1-halo-xcex1,xcex2-saturated carbonyl compound having an absolute configuration of S form at the xcex1-position. The present invention has been accomplished based on these findings.
More specifically, the present invention relates to the following embodiments:
[1] a method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound from an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond by reducing the xcex1,xcex2-carbon-carbon double bond using a microorganism belonging to any one of the genera Acetobacter, Actinomyces, Acinetobacter, Agrobacterium, Aeromonas, Alcaligenes, Arthrobacter, Azotobacter, Bacillus, Brevibacterium, Burkholderia, Cellulomonas, Corynebacterium, Enterobacter, Enterococcus, Escherichia, Flavobacterium, Gluconobacter, Halobacteium, Halococccus, Klebsiella, Lactobacillus, Microbacterium, Micrococcus, Micropolyspora, Mycobacterium, Nocardia, Pseudomonas, Pseudonocardia, Rhodococcus, Rhodobacter, Serratia, Staphylococcus, Streptococcus, Streptomyces and Xanthomonas, or a microbial product thereof;
[2] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1], wherein the xcex1,xcex2-carbon-carbon double bond of the xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond is reduced using a microorganism belonging to the genus Pseudomonas or a microbial product thereof;
[3] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1], wherein the xcex1,xcex2-carbon-carbon double bond of the xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond is reduced using a microorganism belonging to the genus Burkholderia or a microbial product thereof;
[4] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [2], wherein the microorganism belonging to the genus Pseudomonas is Pseudomonas sp. SD810;
[5] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [2], wherein the microorganism belonging to the genus Pseudomonas is Pseudomonas sp. SD811;
[6] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [2], wherein the microorganism belonging to the genus Pseudomonas is Pseudomonas sp. SD812;
[7] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [3], wherein the microorganism belonging to the genus Burkholderia is Burkholderia sp. SD816;
[8] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1] to [7], wherein an S-form compound chiral at the xcex1-position is produced by the reduction of the carbonxe2x80x94carbon double bond;
[9] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1] to [8], wherein the a-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond is a compound represented by the following formula (1): 
wherein R1 represents a halogen atom, R2 and R3 each independently represents a hydrogen atom, a halogen atom, a linear or branched aliphatic hydrocarbon group having from 1 to 6 carbon atoms, a linear or branched alkoxy group having from 1 to 6 carbon atoms, a hydroxyl group, a carboxyl group, an aromatic group which may be substituted, or a nitrogen-, oxygen- or sulfur-containing heterocyclic group, and R4 represents a hydroxyl group, a linear or branched alkoxy group having from 1 to 3 carbon atoms or a primary, secondary or tertiary amino group, and the xcex1-halo-xcex1,xcex2-saturated carbonyl compound is a compound represented by the following formula (2): 
wherein R1 to R4 have the same meanings as defined above;
[10] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [9], wherein the compound represented by formula (1) is an xcex1-haloacrylic acid and the compound represented by formula (2) is an xcex1-halopropionic acid having an absolute S form configuration;
[11] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [10], wherein the halogen atom is a bromine atom;
[12] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [10], wherein the halogen atom is a chlorine atom;
[13] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1] to [12], wherein the microbial product of a microorganism is a microorganism culture, a microbial extract, a microbial cell suspension or a microbial cell fixed to a support;
[14] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1] to [13], wherein the microorganism used is varied not to decompose the xcex1-halo-xcex1,xcex2-saturated carbonyl compound produced, thereby increasing the amount of the product accumulated;
[15] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [1] to [14], wherein the xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond and a compound capable of being oxidized by the microorganism used are present together in the reaction system and thereby the reaction continuing time is prolonged;
[16] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [15], wherein the compound capable of being oxidized by the microorganism used is a sugar having from 3 to 6 carbon atoms;
[17] the method of producing an xcex1-halo-xcex1,xcex2-saturated carbonyl compound as described in [15], wherein the compound capable of being oxidized by the microorganism used is an organic acid having from 2 to 4 carbon atoms;
[18] Pseudomonas sp. SD810 and mutants thereof having an activity of reducing the xcex1,xcex2-carbon-carbon double bond of an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond;
[19] Pseudomonas sp. SD811 and mutants thereof having an activity of reducing the xcex1,xcex2-carbon-carbon double bond of an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond;
[20] Pseudomonas sp. SD812 and mutants thereof having an activity of reducing the xcex1,xcex2-carbon-carbon double bond of an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond;
[21] Burkholderia sp. SD816 and mutants thereof having an activity of reducing the xcex1,xcex2-carbon-carbon double bond of an xcex1-halocarbonyl compound having an xcex1,xcex2-carbon-carbon double bond;
[22] a microbial product containing the microorganism described in [12] to [21]; and
[23] the microbial product as described in [22], which is a microbial culture, a microbial extract, a microbial cell suspension or a microbial cell fixed to a support.